Fig. 6
From: Development and in vitro characterization of humanized antibodies for blocking human TRPM4 channel
Effect of Ab6 (M4H) on cultured human microvascular endothelial cells (HMVECs). (A) Current-voltage relationship of HMVECs that were cultured with control human IgG or Ab6 (M4H) (10 µg/ml) for 30 min before voltage ramps from – 100 to 100 mV were applied. N = 7 cells for both groups. (B) Summary of currents at – 80 or 80 mV from A. (C) Current-voltage relationship of HMVECs that were first cultured with control human IgG (10 µg/ml) for 30 min. Followed by ATP depletion by solution containing 5 mM NaN3 and 10 mM 2-DG applied around 10 μm away from the recording cells. Voltage ramps from – 100 to 100 mV were applied at baseline 0 min and 7 min after ATP depletion treatment. N = 7 cells for both groups. (D) Summary of currents at – 80 or 80 mV from C. (E) Voltage ramps from – 100 to 100 mV at baseline 0 min and 7 min after ATP depletion were applied in cells treated with Ab6 (M4H) (10 µg/ml). N = 7 cells for both groups. (F) Summary of currents at – 80 or 80 mV from E. (G) Sample images of HMVECs treated with control human IgG or Ab6 (M4H) (10 µg/ml) before and after 7-min ATP depletion. Scale bars: 10 μm. (H) Comparison of membrane capacitance (Cm) changes for HMVECs treated with control human IgG and Ab6 (M4H) under ATP depletion induced by 5-mM NaN3 and 10-mM 2-DG for 10 min. In (B, D, F), statistical analysis was performed by Student’s t test and in H by two-way ANOVA with the Bonferroni post hoc test. **P < 0.01, ***P < 0.001, #P < 0.0001.
